wrote the paper. Uncropped scans of the blots shown in the main figures are provided in supplementary Figs. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. COVID-19 is an emerging, rapidly evolving situation. To determine the molecular basis for HIRA homooligomerization, we determined the crystal structure of the C-terminal half of HIRA (aa 644–1017) at 2.4 Å resolution (Table Crystal structure of HIRA(644–1017) reveals a homotrimer. Copyright © 2020, American Association for the Advancement of Science. Having shown that HIRA is able to homooligomerize in vitro as in cells, we examined if Δ759–782 deletion and W799A–D800A double amino acid mutants also exhibit a reduction in the formation of homooligomer in vitro. Furthermore, these three residues are thought to participate in the regulation of histone-histone interaction stability 25 . Winn, M. D. et al. The histone H3 coding sequence probes were hybridized with the green fluorescent dye Fluorescein, and the histone H3-H1 intergenic sequence probes were hybridized with either the far-red fluorescent dye Quasar® 670 Dye (‘Long’, ‘Short Downstream’) or the red fluorescent duy CAL Fluor Red 590® (‘Short Upstream’). Cell infection of HeLa B (from Sandrine Middendorf) and HeLa H3.3-SNAP-HA (from Lars Jansen)Human U2OS (from Jiri Bartek) and HeLa cells were transfected using lipofectamine 2000 and lipofectamine RNAIMAX (Invitrogen) for plasmids and siRNAs, respectively. Dynamics of histone H3 deposition in vivo reveal a nucleosome gap-filling mechanism for H3.3 to maintain chromatin integrity. Im, J. S. et al. and an American Heart Association predoctoral fellowship 12PRE12030157 to M.D.R. Overview of the CCP4 suite and current developments. Lee, J. S. & Zhang, Z. O-linked N-acetylglucosamine transferase (OGT) interacts with the histone chaperone HIRA complex and regulates nucleosome assembly and cellular senescence. and Platform for Drug Discovery, Informatics and Structural Life Science, and Basis for Supporting Innovative Drug Discovery and Life Science Research by AMED (Japan) to T.S. and T.S. HighWire The cells were incubated with buffer A containing 1 mM magnesium chloride, 0.4 mM ATP, 0.02% (w/v) 3-[(3-cholamidopropyl)-dimethylammonio]-propanesulfonate (CHAPS), and 1 U/ml Benozase nuclease (Merck) at 277 K for 30 min. Köhler, A., Schmidt-Zachmann, M. S. & Franke, W. W. AND-1, a natural chimeric DNA-binding protein, combines an HMG-box with regulatory WD-repeats. Eukaryotic histone H3-H4 tetramers contain a putative copper (Cu 2+) binding site at the H3-H3' dimerization interface with unknown function.The coincident emergence of eukaryotes with global oxygenation, which challenged cellular copper utilization, raised the possibility that histones may function in cellular copper homeostasis. You can also search for this author in The canonical histone H3 proteins are incorporated into chromatin via the histone chaperone CAF-1 (chromatin assembly factor 1) at S phase in a replication-dependent manner (1). However, biochemical analysis showed that both mutants eluted entirely in the void volume of a size-exclusion column (Supplementary Fig. De Lucia, F. et al. & Ahmad, K. Nucleosome-depleted chromatin gaps recruit assembly factors for the H3.3 histone variant. Primary antibodies were detected using horse-radish-peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories) and SuperSignal enhanced chemiluminescent detection kit (Pierce). Tagami, H., Ray-Gallet, D., Almouzni, G. & Nakatani, Y. Histone H3.1 and H3.3 complexes mediate nucleosome assembly pathways dependent or independent of DNA synthesis. For the washes 300 mM NaCl was used.For western blot analysis, extracts or immunoprecipitated proteins were run on NuPAGE bis–tris 4–12% gels in MOPS buffer (Invitrogen) and transferred to nitrocellulose membrane (Protran). Identification of an ubinuclein 1 region required for stability and function of the human HIRA/UBN1/CABIN1/ASF1a histone H3.3 chaperone complex.

Tang, Y. et al. Daganzo, S. M. et al. You can also search for this author in generated both HeLa and HeLa H3.3-SNAP-HA cell lines HIRA knockout. You can also search for this author in Transcription factor EKLF (KLF1) recruitment of the histone chaperone HIRA is essential for beta-globin gene expression. Before X-ray diffraction experiments, crystals were flash-frozen using liquid nitrogen. Buschbeck, M. & Hake, S. B. Variants of core histones and their roles in cell fate decisions, development and cancer. Banumathy, G. et al. Tang, Y. et al. Rai, T. S. et al. and R.M. We thank H. Okumura (JASRI) for technical help in the use of beamline BL41XU, T. Ota (KEK) for technical assistance and I. Simeonova for statistical analysis help.Present address: Graduate School of Life Sciences, Tohoku University, 2-1-1, Katahira Aoba-ku, Sendai, Miyagi, 980-8577, JapanThese authors contributed equally: Dominique Ray-Gallet, M. Daniel Ricketts, Yukari Sato.Institut Curie, PSL Research University, CNRS, UMR3664, Equipe Labellisée Ligue contre le Cancer, F-75005, Paris, FranceDominique Ray-Gallet, Ekaterina Boyarchuk & Geneviève AlmouzniSorbonne Université, Institut Curie, CNRS, UMR3664, F-75005, Paris, FranceDominique Ray-Gallet, Ekaterina Boyarchuk & Geneviève AlmouzniDepartment of Biochemistry and Biophysics, Graduate Group in Biochemistry and Molecular Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, 19104, USAStructural Biology Research Center (SBRC), Institute of Materials Structure Science (IMSS), High Energy Accelerator Research Organization (KEK), 1-1 Oho, Tsukuba, Ibaraki, 305-0801, JapanDepartment of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, 19104, USADepartment of Materials Structure Science, School of High Energy Accelerator Science, The Graduate University for Advanced Studies (Soken-dai), 1–1 Oho, Tsukuba, Ibaraki, 305–0801, JapanYou can also search for this author in